Abstract

Methanosaeta strains are frequently involved in the granule formation during methanogenic wastewater treatment. To investigate the impact of Methanosaeta on granulation and performance of upflow anaerobic sludge blanket (UASB) reactors, three 1-L working volume reactors noted as R1, R2, and R3 were operated fed with a synthetic wastewater containing sodium acetate and glucose. R1 was inoculated with 1-L activated sludge, while R2 and R3 were inoculated with 200-mL concentrated pre-grown Methanosaeta harundinacea 6Ac culture and 800mL of activated sludge. Additionally, R3 was daily dosed with 0.5mL/L of acetyl ether extract of 6Ac spent culture containing its quorum sensing signal carboxyl acyl homoserine lactone (AHL). Compared to R1, R2 and R3 had a higher and more constant chemical oxygen demand (COD) removal efficiency and alkaline pH (8.2) during the granulation phase, particularly, R3 maintained approximately 90% COD removal. Moreover, R3 formed the best granules, and microscopic images showed fluorescent Methanosaeta-like filaments dominating in the R3 granules, but rod cells dominating in the R2 granules. Analysis of 16S rRNA gene libraries showed increased diversity of methanogen species like Methanosarcina and Methanospirillum in R2 and R3, and increased bacteria diversity in R3 that included the syntrophic propionate degrader Syntrophobacter. Quantitative PCR determined that 6Ac made up more than 22% of the total prokaryotes in R3, but only 3.6% in R2. The carboxyl AHL was detected in R3. This work indicates that AHL-facilitated filaments of Methanosaeta contribute to the granulation and performance of UASB reactors, likely through immobilizing other functional microorganisms.

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